• The Korean Journal of Malacology
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• v.32 no.2
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• pp.133-139
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• 2016

The incubation time required for hatching of O. ocellatus were investigated through the processes of egg and embryonic developments by the dissecting microscopic and visual observations. And differences in ecological characteristics of the plankton mode of life or the benthic mode of life according to total numbers of the suckers on each short arm of the hatched juvenile larvae of O. ocellatus were studied by comparisons with other octopodidae species. Compared with the recent a few results reported by other researchers associated with the incubation time required for hatching by female adult mother of O. minor (73-90 days after spawning at $20.9-21.5^{\circ}C$ ranges), in this study, the incubation time required for hatching by female adult mother of O. ocellatus was 56-57 days after spawning at $11.0-20.4^{\circ}C$. Therefore, the incubation time required for hatching by female adult mother varied with Octopodidae species. In this studies, each ovarian egg laid by a female was connected to an egg string attaching to the surface of the wall or bottom of vacunt shell of Rapana venosa. Egg and embryonic developments of this species were studied in the indoor aquaria, in the specific gravity ranging 1.024-1.025. the hatched juvenile of O. ocellatus is 10.3 mm in the mean total length and 4.5 mm in mantle length, and each of its short arms has 18-20 suckers. The just hatched juvenile larvae of O. ocellatus enter the benthic mode of life (benthic larval stage) after hatching. In particular, regarding differences in ecological characteristics of the mode of life according to total numbers of the suckers, O. vulgaris may not need to have many suckers because they enter the planktonic mode of life after hatching, however O. ocellatus may need to have many suckers, because they should adapt to the benthic mode of life. And also the just hatched juvenile larvae of O. minor (bearing many suckers more than O. ocellatus) enter the benthic mode of life (benthic larval stage) after hatching. Therefore, the total number of the suckers on each short arm of the hatched juvenile larvae can be used for determining whether an octopus species has planktonic larval stages or benthic larval stage (benthic mode of life). In particular, The intracohort cannibalism phenomena appeared at the hatched juvenile larval stage because the larval stage of O. ocellatus and O. minor enter into the benthic larval stage in the early stage, unlike entering into the plaktonic larval stage in other Octopus species such as O. vulgaris: at this time, the early hatched larvae fed the late hatched larvae (they are the same species and almost same ages). Therefore, the intracohort cannibalism pheneomena occur in the just hatched juvenile stage of only O. ocellatus and O. minor.

• Journal of Sericultural and Entomological Science
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• v.10
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• pp.59-62
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• 1969

It is very important physiological and economical problems to hatching within a day by increasing hatching-ratio of silkworm eggs in sericulture, and modified dark incubating method was experimented in Japan. The author studied on the economical problems of that rather than physiological study. Hatching was induced and accelerated by illumination before hatching 5 days which incubated in the dark room after eye spot pigmented eggs emerged 10-20 percentage. It was increased by 80-90％ in hatching-ratio, but the hatching was delayed a day. In autumn silkworm, newly hatched silkworms were hatched silkworms were hatched in the dark room because of high temperature period tough modified dark incubation was passed only 4 days. The author wonder which is more effective between effect of increasing the hatching-ratio and the health of silkworms.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.345-351
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• 2000

Objective: Hatching of the blastocyst from the zona pellucida (ZP) is a key event in mammalian implantation. In vivo, two factors have been identified as possible mediators of hatching: lysis of the ZP by substances elaborated either from the embryo or female reproductive tract and pressure exerted on the zona by expansion of the blastocyst. Two methods of zona manipulation were already in use to enhance the ability of embryos to hatch: mechanical PZD and chemical ZD by acidic Tyrode's solution. But several controversies of each method have been reported. The purpose of this study was to investigate the effect of pronase for mouse embryo hatching. Methods: Mouse embryos were obtained following ovulation induction of $F_1$ animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ${\mu}g/ml$ pronase in Ham's F10 for 72 hrs. Main outcome measures were the rates of partial hatching and completely hatched blastocysts, and cell number of it. Results: In fresh and cryo-thawed group, the rates of completely hatched blastocyst were significantly higher in 5 ${\mu}g/ml$ pronase treatment group than control group. There was no difference in completely hatched blastocyst total cell number between pronase treatment group and control group. This suggest that pronase treatment did not harmful in mouse embryo development. In pronase treatment group, zona pellucida were thinner than control group. Conclusion: The addition of pronase to culture media may accelerate the hatching of embryo. So, enzymatic treatment of the zona may provide a valuable and effective assisted hatching technique for human in-vitro fertilization-embryo transfer.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.4
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• pp.193-200
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• 2017

Objective: This study was conducted to investigate the efficacy of laser-assisted hatching (LAH) and various vitrification times for embryonic development and blastocyst cell numbers. Methods: First, 2-cell and 8-cell embryos were collected by flushing out the oviducts. In the control groups, they were vitrified for 8 or 10 minutes without LAH. The LAH groups underwent quarter laser zona thinning-assisted hatching before vitrification (4, 6, and 8 minutes or 4, 7, and 10 minutes, respectively). After incubation, double-immunofluorescence staining was performed. Results: The hatched blastocyst rate 72 hours after the 2-cell embryos were thawed was significantly higher in the 2LAH-ES8 group (33.3%) than in the other groups (p< 0.05). In the control-8 group ($22.1{\pm}4.6$), the cell number of the inner cell mass was higher than in the LAH groups (p< 0.05). The number of trophectoderm cells was higher in the 2LAH-ES6 group ($92.8{\pm}8.9$) than in the others (p< 0.05). The hatched blastocyst rate 48 hours after the 8-cell embryos were thawed was higher in the 8LAH-ES4 group (45.5%) than in the other groups, but not significantly. The inner cell mass cell number was highest in the 8LAH-ES7 group ($19.5{\pm}5.1$, p< 0.05). The number of trophectoderm cells was higher in the 8LAH-ES10 group ($73.2{\pm}12.1$) than in the other groups, but without statistical significance. Conclusion: When LAH was performed, 2-cell embryos with large blastomeres had a lower hatched blastocyst rate when the exposure to vitrification solution was shorter. Conversely, 8-cell embryos with small blastomere had a higher hatched blastocyst rate when the exposure to vitrification solution was shorter.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.2
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• pp.68-74
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• 2014

Objective: In search of an ideal method of assisted hatching (AH), we compared the effects of conventional micropipette-AH and laser-AH on the blastocyst formation rate (BFR) and blastocyst cell numbers. Methods: Four-to five-week-old ICR female mice were paired with male mice after superovulation using Pregnant mare's serum gonadotropin (PMSG) and hCG. The two-cell embryos were flushed from the oviducts of female mice. The retrieved two-cell embryos underwent one of five AH procedures: single mechanical assisted hatching (sMAH); cross mechanical assisted hatching (cMAH); single laser assisted hatching (sLAH); quarter laser assisted hatching (qLAH); and quarter laser zona thinning assisted hatching (qLZT-AH). After 72 hours incubation, double immunofluorescence staining was performed. Results: Following a 72 hours incubation, a higher hatching BFR was observed in the control, sMAH, cMAH, and sLAH groups, compared to those in the qLAH and qLZT-AH groups (p<0.05). The hatched BFR was significantly higher in the qLAH and qLZT-AH groups than in the others (p<0.05 for each group). The inner cell mass (ICM) was higher in the control and sMAH group (p<0.05). The trophectoderm cell number was higher in the cMAH and qLAH groups (p<0.05). Conclusion: Our results showed that the hatched BFR was higher in groups exposed the the qLAH and qLZT-AH methods compared to groups exposed to other AH methods. In the qLAH group, although the total cell number was significantly higher than in controls, the ICM ratio was significantly lower in than controls.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.3
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• pp.152-156
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• 2016

Objective: We studied the effect of the site of laser zona opening on the complete hatching of mouse blastocysts and the cell numbers of the completely hatched blastocysts. Methods: Mouse blastocysts were randomly allocated to the inner cell mass (ICM) group (zona opening performed at the site of the ICM, n=125), the trophectoderm (TE) group (zona opening performed opposite to the ICM, n=125) and the control group (no zona opening, n=125). Results: The rate of complete hatching of the blastocysts was not significantly different in the ICM and the TE group (84.8% vs 80.8%, respectively; p=0.402), but was significantly lower in the control group (51.2%, p<0.001). The cell numbers in the completely hatched blastocysts were comparable in the control group, the ICM group, and the TE group ($69{\pm}19.3$, $74{\pm}15.7$, and $71{\pm}16.8$, respectively; p=0.680). Conclusion: These findings indicate that the site of laser zona opening did not influence the rate of complete hatching of mouse blastocysts or their cell numbers.

• Development and Reproduction
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• v.17 no.3
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• pp.247-255
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• 2013

The gonadsomatic index (GSI) of mottled skate was the highest in April, GSI and HSI showed a reverse phase for its reproductive cycle. The fish had one pair of egg capsules, having 1 to 7 fertilized eggs, and spawned all the year round. When surveying the reproductive characteristics of females over 63 cm in disc width, we found the spawning peak was between April to June, and the appearance ratio of egg capsules was the highest in May (32.1%). The eggs were hatched at $8^{\circ}C$, $13^{\circ}C$, $18^{\circ}C$, water temperature (12.8 to $24.2^{\circ}C$), and the best hatching temperature was $18^{\circ}C$. The number of fish hatched was 4 to 5 fish/egg capsules, and the hatching rate was 100%. The sex ratios of hatching larvae were 45.5% female and 54.5% male. Therefore this study will provide fundamental data and information for artificial reproduction of the mottled skate.

• Fisheries and Aquatic Sciences
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• v.8 no.4
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• pp.228-234
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• 2005

Despite the relatively high production of fingerlings of the olive flounder Paralichthys olivaceus, its larval rearing in terms of first feeding has not been fully analyzed. We evaluated the variations of amino acids and fatty acids of starved larvae over 96 hr after hatching. We also investigated depletion of the yolk and oil globule of starved larvae and those fed the rotifer Brachionus plicatilis. In addition, the optimum size of the rotifers according to the mouth size of the larvae, and the point of no return with delay of the first feeding, were also examined. The amino acids in the egg decreased abruptly during embryo development. At 48 to 72 hr after hatching, the amino acids of starved larvae decreased by $30-40\%$ from the level in newly hatched larvae. The concentrations of fatty acids in newly hatched larvae were lower than those of floating eggs and dropped sharply at 48 hr after hatching, when the yolk disappeared. The starved larvae depleted their yolksacs and oil globules earlier than the fed larvae did. At 84 hr after hatching, rotifers were detected for the first time in the guts of the larvae, which were about 3 mm in total length. The point of no return appeared to be close to the fourth day from the first feeding. For a high survival rate of P. olivaceus larvae, the first feeding should occur before the third day after hatching.

• Journal of Aquaculture
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• v.12 no.1
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• pp.25-30
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• 1999

This study was carried out to produce the seedling of the larvae and juveniles of pale chup, Zacco platypus (Cyprinidae) in terms of artificial hatching, feeding behavior and growth rates. The total lengths of the newly hatched larvae were 7.6~8.2mm (mean: 7.97mm). The hatched larvae rose to the surface 3 days after hatching. The larvae were fed Daphnia, rotifer, Artemia and powdered feed. Hatching experiments were conducted in jar incubators under the different water temperature conditions, $20~32^{\circ}C$. The highest hatching rate was at $26^{\circ}C$ after 47 hours. Jar incubators was the highest hatching rate (89%), and cage and Californian incubators were useful and relatively high hatching rates (>89%). The size of the larvae 50 days after hatching were 30.0mm (25.6~32.0mm). As the water temperatures was increased, the growth rate of the larvae was also increased. Especially, the growth rates were favorable over $26^{\circ}C$, but survival was the worst at $32^{\circ}C$. This species was quick to accept assorted feed after hatching. The assorted feeds for flounder and rockfish better than that of carp in reference to growth rate.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.131-137
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• 1997

This study was carried out to investigate the in vivo development rates of vitrified-thawed mouse expanding, hatching and hatched blastoc ysts(BL). In vitro fertilization produced blastocysts were vitrified in EFS40(40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). Expanding a and hatching blastocysts were equilibrated in 20% ethylene glyco](EG) for 5 min. before exposure to EFS40 at 25°C for 1 min., they were then vitrified in liquid nitrogen. Hatched blastocysts which cultured in m-CR1 medium supple mented 0.4% bovine serum albumin on day 5. were equilibrated in 10% EG for 5 min. and then vitrified in EFS40 for 30 sec. After thawing, re-expanding blastocysts were transferred to recipients(3 day of pseudopregnant) on one or both uterine horns(6-8 embryos per a horn). Preg¬n nancy rates of recipients and implantation were a assccessed by autopsy on 15 gestation. The res¬u ults obtained in these experiments were summar¬1 ized as follows; 1) The pregnancy and live fetus rates, for vitrified expanding BL(77.8 and 25.0%) and hatching BL(77.8 and 26.4%)n vitro were not significantly difference in those of control BL (66.7 and 42.9%: 83.3 and 40.4%), respectively, 2) in vitro development of vitrified hatched BL was 34.0%. and 3) in vivo developmental rate of vitrified hatched BL was only 33.3%. These results suggested that proposed rapid vitrification p procedures used EFS40 cryoprotectant can be effectively performed in mouse expanding Ihatching blastocysts and that mouse blastocysts a after being hatched from zona pellucida can be successfully cryopreserved.